We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healui OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healui cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healui (AhCPI) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, $Cys^{25},{\;}His^{159},{\;}and{\;}Asn^{175}$. Deduced amino acid sequence analysis indicates that AhCPI belong to ERFNIN subfamily of C 1 peptidases. By Northern blot analysis. no direct correlation was observed between AhCPI mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that AhCPI protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.
Journal of the Korean Society of Food Science and Nutrition
/
v.28
no.1
/
pp.225-232
/
1999
Apo E polymorphism(e2, e3, e4) was among the first reported genetic polymorphism that explained part of the normal variation in plasma cholesterol concentrations. Among 62 normolipidemic healthy females, aged 19 up to 22 years, the relative frequencies of E3/3 was 0.806(n=50), E3/2 was 0.081(n=5), E3/4 allele was 0.113(n=7), and no E2/2, E2/4 and E4/4 were found. Based on the five samples of E2 allele, five subjects were randomly selected by E3 and E4 groups for the study of effects of apo E polymorphism on the distribution of serum lipid and amino acids profiles. No differences in the anthro pometric data among apo E isomers were found, otherwise the pulsation was higher in E4 than that in the others. There were no differences in plasma total HDL , HDL3 , HDL2 & LDL cholesterol, and apo A I concentrations. However, phenotype means significantly rank E2>E3>E4 allele in average TG levels(p=0.014), and rank E4>E3>E2 in total cholesterol levels(p=0.011). Atherogenic index(AI) such as lipoproteins was significantly increased in E2 & E4 than that in E3(p=0.045). Subjects with E3/2 allele had significantly higher concentrations of glutamine, phosphoserine and taurine, while subjects with E3/4 allele showed significantly lower concentrations of arginine and am inobutyrate and elevated level of phosphoserine in plasma com pared to those of E3/3 allele. Higher level of plasma taurine in subjects with E3/2 or E3/4 allele appears to be related to the elevated level of plasma total and LDL cholesterol concentrations compared to those of E3/3 allele.
The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.
This study was conducted with adult cockerels to determine whether dietary RNA affects feed intake and renal weight and function, and if the responses are similar to dietary adenine. Chickens were ad libitum fed a RNA diet (100 g/kg) or an adenine diet (9.1 g/kg) for 14 d and catheterized in right jugular vein, hepatic portal vein and both urethers, and saline together with para-amino hippuric acid and sodium thiosulfate was continuously infused into them to evaluate renal functions. Dietary RNA reduced feed intake and body weight, and dietary adenine increased kidney weight expressed as a proportion of body weight (P < 0.05). Feed intake and body weight on the adenine diet and kidney weight on the RNA diet showed similar though non significant tendencies. No calculi were detected in the kidney in chickens fed either the RNA or adenine diets. Plasma inorganic phosphate (IP), Ca and 1,25 $(OH)_2$ vitamin $D_3$ concentrations were increased by dietary RNA and adenine, although the increases of IP and Ca in adenine-fed chickens were not significant. Uric acid and urea concentrations in the blood plasma were unaffected by dietary RNA or adenine. Both dietary RNA and adenine increased renal blood flow rates 3.5-3.7 fold, renal plasma flow rates 3.4-3.7 fold and glomerular filtration rates (GFR) 2.9-3.0 fold (p < 0.01). Clearance of urea, IP and Ca were also enhanced by dietary RNA, but not by dietary adenine. However, neither RNA nor adenine affected uric acid clearance. Only IP clearance was significantly augmented at the glomerular level by dietary RNA (p < 0.05). Glomerular filtration of uric acid, urea, IP and Ca and reabsorption of urea, IP and Ca at the renal tubule were increased by dietary RNA and adenine (p < 0.05), whereas tubular secretion of uric acid was decreased by both dietary treatments. It is concluded that dietary adenine is effective in changing renal function and P and Ca metabolism in chickens.
L-carnitine promotes mitochondrial ${\beta}$-oxidation of long chain fatty acids and their subsequent transport across the inner mitochondrial membrane. Although the role of L-carnitine in fatty acid metabolism has been extensively studied, its role in live performance and carcass responses of commercial broilers is less understood. The objective of this research was to determine if Lcarnitine fed at various levels in diets differing in CP and amino acids impacted on live performance and carcass characteristics of commercial broilers. Two floor pen experiments were conducted to assess the effect of dietary L-carnitine in grower diets. In Exp. 1, Ross${\times}$Hubbard Ultra Yield broilers were placed in 48 floor pens (12 birds/pen) and fed common diets to d 14. A two (0 or 50 ppm Lcarnitine) by three (173, 187, and 202 g/kg CP) factorial arrangement of treatments was employed from 15 to 35 d of age (8 replications/treatment). An interaction (p<0.05) in carcass yield indicated that increasing CP (187 g/kg) resulted in improved yield in the presence of L-carnitine. Increasing CP from 173 to 202 g/kg increased (p<0.05) BW gain and decreased (p<0.05) feed conversion and percentage abdominal fat. Feeding dietary L-carnitine increased back-half carcass yield which was attributable to an increase (p<0.05) in thigh, but not drumstick, yield relative to carcass. In Exp. 2, $Ross{\times}Ross$ 708 broilers were fed common diets until 29 d. From 30 to 42 d of age, birds were fed one of seven diets: i) 200 g/kg CP, 0 ppm L-carnitine; ii) 200 g/kg CP, 40 ppm L-carnitine; iii) 180 g/kg CP, 0 ppm L-carnitine; iv) 180 g/kg CP, 10 ppm L-carnitine; v) 180 g/kg CP, 20 ppm L-carnitine; vi) 180 g/kg CP, 30 ppm L-carnitine; and vii) 180 g/kg CP, 40 ppm L-carnitine (6 replications of 12 birds each). BW gain, feed conversion, mortality (30 to 42 d), and carcass traits (42 d) were measured on all birds by pen. There were no treatment differences (p<0.05). However, the addition of 40 ppm L-carnitine in the 200 g CP/kg diet increased (p = 0.06) thigh yields relative to BW in comparison to birds fed diets without L-carnitine, which was further confirmed via a contrast analysis (0 vs. 40 ppm L-carnitine in the 200 and 180 g CP/kg diets; p<0.05). These results indicated that dietary L-carnitine may heighten metabolism in dark meat of commercial broilers resulting in increased relative thigh tissue accretion without compromising breast accretion.
Due to the recent economic development, the diet style has become more and more westernized in Korea, which increased the concern of our well-beings. Our well-beings are also associated with the gut microbiota which vary depending on the dietary intake. In this study, we compared gut microbiome shifted by two diets: high-fat diets (HFD) and low-fiber diet (LFD) based on 16S rRNA gene sequences using MiSeq. Compared to the control diet, LFD and HFD treatments significantly decreased species richness, while there was no difference in species evenness. Both diet treatments significantly increased the relative abundance of the Proteobacteria (p<0.05), especially the genus Sutterella. Bacteroidetes was significantly decreased in HFD groups, where the family S24-7 was decreased most. On the other hand, significant difference between HFD and LFD was seen among Firmicutes, where the abundance of family Lachnospiraceae was lower in LFD groups (p<0.05). PICRUSt-based metabolic difference analyses showed LFD treatment significantly decreased metabolisms of amino acid, carbohydrate and methane (p<0.01). In contrast, HFD significantly increased amino acid metabolism (p<0.05). Glycan biosynthesis and metabolism were significantly increased in both treatment groups (p<0.01). Our results suggest that long-term unbalanced dietary intakes induce gut dysbiosis, leading to metabolic and colonic disorders.
Insulin-like growth factor-I(IGF-I) has significant insulin-like anabolic effects which include the stimulation of glucose and amino acid uptake, as well as protein and glycogen synthesis. IGFs exist in serum and other biological fluids as complexes bound to a family of structurally related insulin-like growth factor binding proteins(IGFBPs). Six human IGFBPs can modulate the effects of IGFs on target tissues by several mechanisms, including altering the serum's half-life and the transcapillary transport of IGFs, as well as changing the availability of IGFs to specific cell surface receptors. Human fibroblasts secrete IGFBPs that can modify IGF-I action. Previous to our study using either Northern blotting, and Western blotting have shown that fibroblasts express mRNA IGFBP-3, -4, and -5, and synthesize these proteins. In addition, fibroblast cell lysates revealed that the IGFBP-3 was most abundant. For these reasons, we undertook to gain further insight into the effects of high and low glucose incubation condition on metabolism and IGFBP-3 expression. In results of metabolites and IGFBP-3 expression in GM10 cells cultivated with various glucose concentration, the consumption of glucose and accumulation of triglyceride were increased in condition of high glucose, and total protein level was decreased. in the course of time. After 5 days incubation, levels of free amino acid in medium containing glucose of high concentration glucose were higher than in conditions of low glucose. Although the levels of IGFBP-3 protein and mRNA levels were increased in low glucose, and IGFBP-3 was not affected by any pretense. Taken together, we suggest that the study of growth factors, like IGFs, might be a possible model of diabetes militus in cell, although the results in cell models were not in accord with in vivo.
The aim of this study was to examine the effects of Capsosiphon fulvescens extract (CFE) on levels of essential amino acids in serum and the rate of protein digestion. CFE contains approximately 68% carbohydrate as fiber, 17% crude ash, and 14% crude protein. Experimental rats were divided two groups, one group receiving a normal diet and the other a normal diet containing 5% CFE. To analyze the absorption of essential amino acids into the hepatic portal vein, blood from the hepatic portal vein was collected between 10 a.m. and 5 p.m. for 10 days. In vitro protein digestion was decreased in the group fed 5% CFE; these data indicated that CFE affected protease activity. We measured the absorption of essential amino acids in the serum of the hepatic portal vein, at 30-, 60-, 90-, and 120 minutes after feeding. Although there was no difference in the concentration of total essential amino acids between the two groups, the 5% CFE-fed animals had a decreased rate of absorption. Absorption of Lys and Thr into the hepatic portal vein was lower in the CFE-fed group than in the control group. The rate of absorption of Met was delayed nearly 50% in the CFE-fed group compared to the control group. On the other hand, the rate of absorption of Leu, Ile, and Val was increased; Phe showed the same. Therefore, we suggest that CFE could affect protein metabolism by increasing or decreasing the absorption rate of essential amino acids.
Hwang Byung Kook;Ibenthal Wolf-Dieter;Heitefuss Rudolf
Korean Journal Plant Pathology
/
v.2
no.1
/
pp.22-30
/
1986
The effect of powdery mildew infection on the $^{14}CO_2$ assimilation and metabolism of $^{14}C-$assimilates was studied with spring barley cultivars, susceptible Peruvian and adult-plant resistant Asse at the four-leaf stage. No consistent differences between Peruvian and Asse were revealed in $^{14}CO_2$ assimilation and metabolism of $^{14}C-$assimilates in healthy whole plants. In the two cultivars, $^{14}CO_2$ assimilation and translocation of assimilates decreased as the number of infected leaves increased. Despite the same infection intensity, $^{14}CO_2$ assimilation was less inhibited in Asse than Peruvian. Infection reduced the fixation of $^{14}CO_2$ in noninfected fourth leaves of Peruvian more severely than that of Asse. Infection of the lower 3 leaves also inhibited the incorporation of 14 C into carbohydrates such as fructose and glucose in noninfected fourth leaves and their translocation into leaf sheathes, the inhibitions being greater in Peruvian than Asse. In the infected third leaves, there was a reduction of 14 C-activity in carbohydrates, more $^{14}C-$labeled fructose and glucose being retained in Peruvian. The stimulation of $^{14}C-$organic acid synthesis in all plant organs was more pronounced in Peruvian than Asse. Powdery mildew markedly increased the incorporation of $^{14}C$ into amino acids in infected third and noninfected fourth leaves, but reduced their translocation to the leaf sheathes. A greater rise of $^{14}C-$ activity in some amino acids in the two leaves was found in Peruvian than Asse.
Objective: This study was conducted to evaluate the effects of inclusion level and amino acid (AA) supplementation on energy values of soybean oil (SO) as determined by difference method or regression method when fed to growing pigs. Methods: Thirty-six barrows (initial body weight: 28.0±1.3 kg) were randomly assigned to one of 6 dietary treatments, which included 2 control diets formulated using a basal diet with or without AA supplementation, and 4 experimental diets with 5% or 10% SO addition in the 2 control diets, respectively. All pigs were individually housed in metabolism crates for 19 d, and during the last 5 d, total urine and feces production were collected. The nutrient digestibility in diets and the digestible energy (DE) and metabolizable energy (ME) values of SO were determined using the difference method and the regression method, respectively. Results: Our results showed that there were no interaction effects (p>0.05) between AA supplementation and SO inclusion levels on energy values of SO and dietary nutrient digestibility. The DE and ME values of SO determined by the difference method were not affected (p>0.05) by AA supplementation, however, the ME value of SO increased (p<0.05) as the inclusion level of SO increased. Moreover, the energy values of SO determined using the regression method were close to those determined using difference method with 10% SO inclusion, but were greater than those obtained using difference method with 5% SO inclusion. Conclusion: We concluded that the DE and ME values of SO increased with the inclusion level but were not affected by AA supplementation in the range of 0% to 10%. The difference method can substitute for the regression method to determine the DE and ME values of SO when the inclusion level is 10%, but not at 5% inclusion level.
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